{"created":"2023-07-27T06:58:27.453239+00:00","id":56894,"links":{},"metadata":{"_buckets":{"deposit":"a853ae9d-0a84-4a60-86ea-485890d7aebc"},"_deposit":{"created_by":18,"id":"56894","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"56894"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00056894","sets":["2812:2813:2829"]},"author_link":["1725"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2006-07-10","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"2p.","bibliographicVolumeNumber":"2003 – 2004","bibliographic_titles":[{"bibliographic_title":"平成16(2004)年度 科学研究費補助金 基盤研究(C) 研究成果報告書概要"},{"bibliographic_title":"2004 Fiscal Year Final Research Report Summary","bibliographic_titleLang":"en"}]}]},"item_9_creator_33":{"attribute_name":"著者別表示","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{}],"nameIdentifiers":[{},{}]}]},"item_9_description_21":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"本研究では、間期の細胞においてGRASP65の277番目のセリン(S277)がリン酸化されていること、また、このリン酸化がEGFなどの増殖刺激で増大することを明らかにした。また、EGFによる増殖シグナルが、ERKを活性化させること、また活性化ERKがS277を直接リン酸化することを明らかにした。さらに、S277が、M期において顕著にリン酸化されることを見いだし、その分子機構の解析を行った。S277付近のアミノ酸配列は(PGSPG)であって、ほ乳類のGRASP65で良く保存されていた。この部位は、cdk1/cyclinBのリン酸化酵素の認識配列に合致していた。M期の細胞質抽出液でGRASP65を処理するとS277は顕著にリン酸化され、これはcdkの特異的阻害剤であるroscovitinで阻害された。一方、MEKの阻害剤であるU0126存在下で調整し、ERKを不活化したM期の細胞質抽出液でもS277はリン酸化された。以上のことから、M期でのS277のリン酸化は、cdk1/cyclinBによっておこり、ERKは関与しないことが強く示唆された。驚いたことに、N末がミリスチン酸修飾されないGRASP65(Δm-GRASP65)を精製しG2期の細胞の細胞質に導入すると、M期への進行が顕著に阻害されることを見いだした。M期への進行阻害は、導入するΔm-GRASP65のS277をアラニンに変異させるともはや観察されなかった。従って、Δm-GRASP65は、S277部位において何らかの細胞質因子と相互作用して細胞周期の進行を阻害していることが示唆された。さらに、リン酸化されたS277部位にPlk1が特異的に結合すること、また非リン酸化状態のS277部位に特異的に結合するタンパク質が存在することも見いだした。","subitem_description_type":"Abstract"},{"subitem_description":"We have shown that a 277th amino acid residue of GRASP65 (S277) is phosphorylated in interphase cells and the phosphorylation signal is markedly enhanced by the growth factor treatment including EGF. ERK is activated by the EGF induced growth factor signal and the activated ERK phosphorylates S277 directly. We further found that S277 is heavily phosphorylated during M phase and analyzed the molecular mechanism for this up regulation of the phosphorylation. The amino acid sequence around S277 (PGSPG) is well conserved among mammalian CTRASP65 homologues. This sequence is well fitted with the target sequence of cdk1/cyclinB. S277 was strongly phosphorylated by a cytoplasmic extract of M phase cells and this was completely inhibited by roscovitine, a cdk specific inhibitor. S277 was also phosphorylated by an ERK inactive cytoplasmic extract of M phase cells that was prepared in the presence of U0126, a MEK inhibitor. These results strongly suggested that cdk1/cyclinB, and not ERK, is responsible for the phosphorylation of S277 in M phase. Surprisingly, the mitotic entry was strongly inhibited by the microinjection of purified GRASP65 without N-terminal myristoylation (Δm-GRASP65). This was not observed by the microinjection of Δm-GRASP65 in which S277 was changed with alanine. These results suggested that Δm-GRASP65 interact with some cytoplasmic factors and inhibits the mitotic entry. We have found that Plk1 specifically binds to phosphorylated S277 region of GRASP65 and there are some cytoplasmic factors that bind to unphosphorylated S277 region of GRASP65.","subitem_description_type":"Abstract"}]},"item_9_description_22":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"研究課題/領域番号:15570156, 研究期間(年度):2003 – 2004","subitem_description_type":"Other"},{"subitem_description":"出典：「ゴルジ体による細胞機能調節機構の解明」研究成果報告書　課題番号15570156\n（KAKEN：科学研究費助成事業データベース（国立情報学研究所））\n（https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-15570156/155701562004kenkyu_seika_hokoku_gaiyo/)を加工して作成","subitem_description_type":"Other"}]},"item_9_description_5":{"attribute_name":"提供者所属","attribute_value_mlt":[{"subitem_description":"金沢大学自然科学研究科","subitem_description_type":"Other"}]},"item_9_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00063168","subitem_identifier_reg_type":"JaLC"}]},"item_9_relation_28":{"attribute_name":"関連URI","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"https://nrid.nii.ac.jp/ja/search/?kw=50294955"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://nrid.nii.ac.jp/ja/search/?kw=50294955","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-15570156/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-15570156/","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-15570156/155701562004kenkyu_seika_hokoku_gaiyo/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-15570156/155701562004kenkyu_seika_hokoku_gaiyo/","subitem_relation_type_select":"URI"}}]},"item_9_version_type_25":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_ab4af688f83e57aa","subitem_version_type":"AM"}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2021-11-08"}],"displaytype":"detail","filename":"SC-PR-NAKAMURA-N-kaken 2006-2p.pdf","filesize":[{"value":"88.8 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"SC-PR-NAKAMURA-N-kaken 2006-2p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/56894/files/SC-PR-NAKAMURA-N-kaken 2006-2p.pdf"},"version_id":"242a0d14-13e4-402a-b219-b9adb0be53a8"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"ゴルジ体による細胞機能調節機構の解明","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"ゴルジ体による細胞機能調節機構の解明"},{"subitem_title":"EXAMINATION OF REGULATROY MECHANISMS FOR CELL FUNCTIONS BY THE GOLGI APPARATUS","subitem_title_language":"en"}]},"item_type_id":"9","owner":"18","path":["2829"],"pubdate":{"attribute_name":"公開日","attribute_value":"2021-11-08"},"publish_date":"2021-11-08","publish_status":"0","recid":"56894","relation_version_is_last":true,"title":["ゴルジ体による細胞機能調節機構の解明"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-07-27T14:25:58.124354+00:00"}